Preparation method and application of compound plant volatile oil for preventing and treating bacterial enteritis of meat ducks

Abstract

The invention belongs to the technical field of veterinary drugs, discloses a preparation method and application of compound plant volatile oil for preventing and treating bacterial enteritis of meat ducks, and particularly discloses a water-soluble compound plant volatile oil solution. Comprising the following components: cinnamon volatile oil, origanum volatile oil, vitamins, oligosaccharide, a surfactant, a co-emulsifier, a preservative and an antioxidant. The compound plant volatile oil solution disclosed by the invention has a relatively strong in-vitro antibacterial effect, can replace antibiotics such as neomycin sulfate, apramycin and colistin sulfate to effectively prevent and treat enteritis of the meat ducks, has no withdrawal period and no medicine residue, improves the clinical dilute fecal symptom, improves the intestinal health degree and slaughter weight of ducks, ensures the slaughter rate, reduces the feed conversion ratio, and has a good market prospect. The weight of duck gizzards and duck intestines is increased, and the economic benefits of meat duck breeding and slaughtering are improved.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary medicines, and particularly relates to a preparation method and application of a composite plant volatile oil for preventing and treating bacterial enteritis in meat ducks. Background Art

[0002] In recent years, with the diversified development of market demand, the meat duck farming model has rapidly transitioned from individual farming to intensive and large-scale farming. While production capacity has expanded rapidly, although the meat duck marketing cycle has been effectively shortened, high-density farming has also increased the risk of disease transmission. Among them, bacterial enteritis is one of the representative stubborn diseases in meat duck farming.

[0003] Bacterial enteritis in meat ducks often presents with lethargy, loss of appetite, or even complete loss of appetite, loose, poorly formed feces, and large amounts of yellowish-white feces adhering to the perianal area. In severe cases, ducks may die or experience stunted growth. Post-mortem examination reveals scant intestinal contents with a strong fishy odor, and congestion, hemorrhage, or ulcers in the small intestinal mucosa. Failure to promptly intervene with medication can lead to emaciation, increased mortality, and decreased feed conversion, directly impacting farm profitability.

[0004] Bacterial enteritis in meat ducks can be caused by Escherichia coli, Salmonella, Clostridium perfringens, and Riemerella anatipestifer. Antibiotics still dominate the treatment options. Neomycin, apramycin, and colistin sulfate are the mainstays of clinical antibiotic prevention and control for bacterial enteritis in meat ducks. The increased use of antibiotics can lead to increased bacterial resistance, decreased sensitivity to conventional treatments, and decreased efficacy, which in turn increases drug dosage and treatment costs for farmers. Furthermore, antibiotics are slowly metabolized, leaving residues in duck meat, posing a threat to food safety and human health.

[0005] In this context, microecological agents, Chinese herbal medicines, and antibody products for replacing antibiotics in aquaculture have gradually become popular in development. Especially for Chinese herbal medicines, due to their multiple advantages of wide sources, low cost, difficulty in generating drug resistance, and no withdrawal period, they have been widely studied for preventing and treating bacterial enteritis in meat ducks. The traditional Chinese veterinary medicine formula, Baitouweng Koufuye, was earlier used for treating poultry enteritis. This formula includes 150 g of Coptis chinensis, 300 g of Pulsatilla chinensis, 300 g of Cortex Fraxini, and 225 g of Phellodendron amurense. It has the effects of clearing heat and detoxifying, cooling blood and stopping dysentery, and also has a certain antibacterial effect. However, both Coptis chinensis and Pulsatilla chinensis belong to precious medicinal materials. In the current low-profit breeding environment of meat ducks, it is difficult to bear the use cost of this product in actual breeding. Poplar flower oral liquid has also been widely used for bacterial enteritis in meat ducks due to its effect of promoting digestion and stopping dysentery. However, in clinical applications, it often requires a long treatment cycle and has a slow onset, which greatly restricts its clinical application. Under the influence of traditional formulas, more and more Chinese medicine compositions have been developed, verified, and applied to the treatment of clinical bacterial enteritis in meat ducks. For example, patent document CN108815283A discloses a traditional Chinese medicine probiotic compound preparation for preventing and treating Salmonella enteritidis infection in ducklings and its preparation method, including: 5 - 15 parts of Portulaca oleracea, 5 - 8 parts of Glycyrrhiza uralensis, 4 - 5 parts of Platycladus orientalis, 5 - 9 parts of Phellodendron amurense, 1 - 3 parts of Plantago asiatica, 2 - 5 parts of Prunus mume, 1 - 3 parts of Cassia obtusifolia, 2 - 6 parts of Citrus reticulata Blanco, 5 - 8 parts of Magnolia officinalis, and 5 - 6 parts of Endothelium Corneum Gigeriae Galli. The combination of these drugs has the effects of clearing heat and detoxifying, promoting digestion and strengthening the spleen, not only improving the immunity of the duck flock, but also helping the duck flock to absorb nutrients, improving production efficiency, and having a certain effect on preventing and controlling enteritis in ducklings.

[0006] The applicant has found that most of the publicly disclosed effective antibiotic - replacing formulas have certain limitations. Either they have a single function and only have a certain antibacterial activity, or their treatment application effects are limited, or the cost of cycle application is too high. Based on these problems, the applicant attempts to purify and concentrate the active ingredients of plant extracts to achieve high - activity antibacterial and treatment effects on meat duck enteritis. In recent years, plant essential oils have been widely reported for their high antibacterial activity, but their treatment effects on meat duck enteritis are not yet clear. Moreover, the extraction processes of plant essential oils vary widely, often resulting in low extraction efficiency of active ingredients. In addition, plant essential oils have poor water solubility and are difficult to administer through the water line, severely restricting their clinical applications.

[0007] In response to these problems, there is an urgent need to develop a natural, antibiotic - replacing plant essential oil composition that is effective in treating meat duck enteritis. At the same time, explore and optimize the essential oil extraction process, then construct a system for the essential oil, create its water solubility, achieve uniform dispersion and administration through the water line in clinical settings, and finally compound with intestinal repair factors to enhance the astringent effect on meat duck enteritis, promote the duck flock to recover growth performance, which is of great significance for preventing and treating stubborn enteritis in large - scale meat duck farming.

[0008] Therefore, this application is specifically proposed. Summary of the Invention

[0009] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. The present invention provides a preparation method and application of a composite plant volatile oil for preventing and treating bacterial enteritis in meat ducks, which has strong in vitro antibacterial efficacy, can replace antibiotics such as neomycin, apramycin, and colistin to effectively prevent and treat enteritis in meat ducks, has no withdrawal period, no drug residues, can improve the clinical symptom of loose feces, enhance intestinal health, ensure the slaughter rate, reduce the feed-to-meat ratio, increase the yield rate of duck gizzards, and improve the economic benefits of meat duck breeding and slaughtering.

[0010] The purpose of the first aspect of the present invention is to provide a water-soluble composite plant volatile oil solution.

[0011] The purpose of the second aspect of the present invention is to provide a preparation method of the water-soluble composite plant volatile oil solution of the first aspect of the present invention.

[0012] The purpose of the third aspect of the present invention is to provide a natural medicine.

[0013] The purpose of the fourth aspect of the present invention is to provide the application of the water-soluble composite plant volatile oil solution of the first aspect of the present invention and / or the medicine of the third aspect of the present invention.

[0014] The purpose of the fifth aspect of the present invention is to provide a product.

[0015] In order to achieve the above purpose, the technical solutions adopted by the present invention are as follows:

[0016] In the first aspect of the present invention, there is provided a water-soluble composite plant volatile oil solution, which includes the following components: cinnamon volatile oil, oregano volatile oil, vitamins, oligosaccharides, surfactants, co-emulsifiers, preservatives, and antioxidants.

[0017] In some embodiments of the present invention, by mass, the water-soluble composite plant volatile oil solution includes 7-12 parts of cinnamon volatile oil, 6-10 parts of oregano volatile oil, 0.5-1.5 parts of vitamins, 0.01-0.1 parts of oligosaccharides, 30-45 parts of surfactants, 60-85 parts of co-emulsifiers, 0.005-0.4 parts of preservatives, and 0.01-0.5 parts of antioxidants.

[0018] In some preferred embodiments of the present invention, by mass, the water-soluble composite plant volatile oil solution includes 8-10 parts of cinnamon volatile oil, 7-9 parts of oregano volatile oil, 0.5-1 part of vitamins, 0.05-0.1 parts of oligosaccharides, 35-40 parts of surfactants, 70-80 parts of co-emulsifiers, 0.1-0.3 parts of preservatives, and 0.05-0.3 parts of antioxidants.

[0019] In some embodiments of the present invention, the vitamins include at least one of vitamin A, vitamin D3, vitamin E, and vitamin K3.

[0020] In some preferred embodiments of the present invention, the vitamins include vitamin A, vitamin E, and vitamin K3. More preferably, the vitamins include vitamin A or vitamin K3; most preferably, the vitamin is vitamin A.

[0021] In some embodiments of the present invention, the oligosaccharides include at least one of mannan oligosaccharide and xylo-oligosaccharide; preferably, the oligosaccharide is mannan oligosaccharide.

[0022] In some embodiments of the present invention, the surfactants include at least one of Tween-40, Tween-60, and Tween-80; preferably, the surfactant is Tween-80.

[0023] In some embodiments of the present invention, the co-emulsifiers include at least one of ethanol, glycerol, propylene glycol monocaprylate, polyethylene glycol glycerol caprylate / caprate, and diethylene glycol monoethyl ether.

[0024] In some preferred embodiments of the present invention, the co-emulsifiers include at least one of propylene glycol monocaprylate, polyethylene glycol glycerol caprylate / caprate, and diethylene glycol monoethyl ether; more preferably, the co-emulsifiers include at least one of polyethylene glycol glycerol caprylate / caprate and diethylene glycol monoethyl ether; most preferably, the co-emulsifier is a combination of polyethylene glycol glycerol caprylate / caprate and diethylene glycol monoethyl ether.

[0025] In some embodiments of the present invention, the preservatives include at least one of potassium cinnamate, sodium benzoate, potassium sorbate, and ethylparaben.

[0026] In some preferred embodiments of the present invention, the preservatives include at least one of potassium cinnamate, sodium benzoate, and ethylparaben; more preferably, the preservatives include at least one of potassium cinnamate and sodium benzoate; most preferably, the preservative is potassium cinnamate.

[0027] In some embodiments of the present invention, the antioxidants include at least one of butylated hydroxyanisole, dibutylhydroxytoluene, tea polyphenols, and tert-butylhydroquinone.

[0028] In some preferred embodiments of the present invention, the antioxidants include at least one of butylated hydroxyanisole, dibutylhydroxytoluene, and tea polyphenols; more preferably, the antioxidants include at least one of butylated hydroxyanisole and tea polyphenols; most preferably, the antioxidant is a combination of butylated hydroxyanisole and tea polyphenols.

[0029] In some embodiments of the present invention, the water-soluble composite plant volatile oil solution comprises 8-10 parts of cinnamon volatile oil, 7-9 parts of oregano volatile oil, 0.5-1 part of vitamin A, 0.05-0.1 part of mannan oligosaccharide, 35-40 parts of Tween-80, 5-10 parts of polyethylene glycol glycerol caprylate / caprate, 55-75 parts of diethylene glycol monoethyl ether, 0.1-0.3 part of potassium cinnamate, 0.03-0.2 part of butylated hydroxyanisole, and 0.02-0.1 part of tea polyphenols.

[0030] In some embodiments of the present invention, the water-soluble composite plant volatile oil solution further comprises 3-10 parts of water.

[0031] The second aspect of the present invention provides a preparation process for cinnamon volatile oil:

[0032] The dried cinnamon medicinal material is ultrafinely pulverized and sieved through a mesh sieve to obtain a cinnamon pulverized material;

[0033] The cinnamon pulverized material is infiltrated with ethanol to obtain an infiltrated cinnamon medicinal material;

[0034] Then the infiltrated cinnamon medicinal material is subjected to steam explosion to obtain a cinnamon steam explosion treated material;

[0035] The volatile oil of the cinnamon steam explosion treated material is extracted and concentrated by using the supercritical CO2 extraction method to obtain the cinnamon volatile oil.

[0036] In some embodiments of the present invention, the mesh number of the mesh sieve is 20-40 meshes; preferably 30 meshes.

[0037] In some embodiments of the present invention, the concentration of the ethanol is 20-40%, and the water content is controlled at 20%-30% after treatment.

[0038] In some embodiments of the present invention, the pressure of the steam explosion condition is 0.6-1.5 MPa.

[0039] In some preferred embodiments of the present invention, the pressure of the steam explosion condition is 0.7-1.0 MPa.

[0040] In some embodiments of the present invention, the pressure maintenance time of the steam explosion condition is 60-240 s.

[0041] In some preferred embodiments of the present invention, the pressure maintenance time of the steam explosion condition is 80-160 s.

[0042] In some embodiments of the present invention, the explosion time of the steam explosion condition is 0.01 s.

[0043] In some embodiments of the present invention, the extraction pressure of the supercritical CO2 extraction is 15-50 MPa.

[0044] In some preferred embodiments of the present invention, the extraction pressure of the supercritical CO2 extraction is 20 to 35 MPa.

[0045] In some embodiments of the present invention, the extraction temperature of the supercritical CO2 extraction is 30 to 60 °C.

[0046] In some preferred embodiments of the present invention, the extraction temperature of the supercritical CO2 extraction is 35 to 45 °C.

[0047] In some embodiments of the present invention, the extraction time of the supercritical CO2 extraction is 1 to 4 h.

[0048] In some preferred embodiments of the present invention, the extraction time of the supercritical CO2 extraction is 1.5 to 3.5 h.

[0049] In some embodiments of the present invention, the CO2 flow rate of the supercritical CO2 extraction is 5 to 30 L / h.

[0050] In some preferred embodiments of the present invention, the CO2 flow rate of the supercritical CO2 extraction is 10 to 20 L / h.

[0051] In some embodiments of the present invention, the entrainer for the supercritical CO2 extraction includes one of methanol, ethanol, and acetone.

[0052] In some preferred embodiments of the present invention, the entrainer for the supercritical CO2 extraction is ethanol.

[0053] In some embodiments of the present invention, the addition ratio of the entrainer is 1% to 8%.

[0054] In some preferred embodiments of the present invention, the addition ratio of the entrainer is 2% to 6%.

[0055] The third aspect of the present invention provides a preparation process for oregano volatile oil:

[0056] Ultra-finely pulverize the dried oregano medicinal material and pass through a sieve to obtain the pretreated oregano medicinal material;

[0057] Perform ethanol infiltration on the pretreated oregano medicinal material to obtain the infiltrated oregano medicinal material;

[0058] Then perform steam explosion on the infiltrated oregano medicinal material to obtain the oregano steam explosion treatment product;

[0059] Use the supercritical CO2 extraction method to extract and concentrate the volatile oil from the oregano steam explosion treatment product to obtain the oregano volatile oil.

[0060] In some embodiments of the present invention, the mesh number of the mesh sieve is 30 to 50 meshes; preferably 40 meshes.

[0061] In some embodiments of the present invention, the concentration of the ethanol is 20% to 40%, and the water content is controlled to be 20% to 30% after treatment.

[0062] In some embodiments of the present invention, the pressure of the steam explosion condition is 0.4 to 1.2 MPa.

[0063] In some preferred embodiments of the present invention, the pressure of the steam explosion condition is 0.5 to 0.8 MPa.

[0064] In some embodiments of the present invention, the holding time of the steam explosion condition pressure is 60 to 180 s.

[0065] In some preferred embodiments of the present invention, the holding time of the steam explosion condition pressure is 60 to 120 s.

[0066] In some embodiments of the present invention, the explosion time of the steam explosion condition is 0.01 s.

[0067] In some embodiments of the present invention, the extraction pressure of the supercritical CO2 extraction condition is 8 to 40 MPa.

[0068] In some preferred embodiments of the present invention, the extraction pressure of the supercritical CO2 extraction condition is 10 to 25 MPa.

[0069] In some embodiments of the present invention, the extraction temperature of the supercritical CO2 extraction condition is 35 to 50 °C.

[0070] In some preferred embodiments of the present invention, the extraction temperature of the supercritical CO2 extraction condition is 35 to 40 °C.

[0071] In some embodiments of the present invention, the extraction time of the supercritical CO2 extraction condition is 1 to 3 h.

[0072] In some preferred embodiments of the present invention, the extraction time of the supercritical CO2 extraction condition is 1 to 2 h.

[0073] In some embodiments of the present invention, the CO2 flow rate of the supercritical CO2 extraction condition is 1 to 35 L / h.

[0074] In some preferred embodiments of the present invention, the CO2 flow rate of the supercritical CO2 extraction condition is 15 to 25 L / h.

[0075] In some embodiments of the present invention, the supercritical CO2 extraction entrainer includes one of methanol, ethanol, and acetone.

[0076] In some preferred embodiments of the present invention, the supercritical CO2 extraction entrainer is ethanol.

[0077] In some embodiments of the present invention, the addition ratio of the entrainer is 3% - 10%.

[0078] In some preferred embodiments of the present invention, the addition ratio of the entrainer is 5% - 9%.

[0079] The fourth aspect of the present invention provides a method for preparing the water-soluble composite plant volatile oil solution of the first aspect of the present invention, comprising the following steps:

[0080] Stir and mix vitamins, oligosaccharides, preservatives, antioxidants and co-emulsifiers to obtain Solution 1;

[0081] Mix Solution 1 with a surfactant to obtain Solution 2;

[0082] Mix Solution 2 with cinnamon volatile oil and oregano volatile oil and emulsify to obtain the water-soluble composite plant volatile oil solution.

[0083] In some embodiments of the present invention, the emulsification pressure is 25 - 50 MPa.

[0084] In some preferred embodiments of the present invention, the emulsification pressure is 30 - 45 MPa.

[0085] In some embodiments of the present invention, the emulsification temperature is 25°C - 50°C.

[0086] In some preferred embodiments of the present invention, the emulsification temperature is 35 - 45°C.

[0087] In some embodiments of the present invention, the emulsification time is 0.25 - 1.5 h.

[0088] In some preferred embodiments of the present invention, the emulsification time is 0.5 - 1 h.

[0089] The fifth aspect of the present invention provides a drug comprising the water-soluble composite plant volatile oil solution of the first aspect of the present invention.

[0090] In some embodiments of the present invention, the drug comprises pharmaceutically acceptable excipients.

[0091] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of fillers, disintegrants, diluents, dispersants, excipients, stabilizers, lubricants, binders, wetting agents, flavoring agents, suspending agents, solvents, sustained-release agents, emulsifiers, absorption promoters, surfactants, preservatives, pigments, flavors and solvents.

[0092] In some embodiments of the present invention, the filler is selected from starch, sucrose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, glucose, etc.; the binder is selected from cellulose derivatives, alginates, starch, water, dextrin, gelatin, polyvinylpyrrolidone, etc.; the disintegrant is selected from microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linked sodium carboxymethyl cellulose; the lubricant is selected from stearic acid, polyethylene glycol, calcium carbonate, sodium bicarbonate, colloidal silicon dioxide, talc, magnesium stearate; the suspending agent is selected from colloidal silicon dioxide, beeswax, cellulose, solid polyethylene glycol; the wetting agent is selected from glycerol, Tween-80, hydrogenated castor oil or lecithin; the solvent is selected from ethanol, liquid polyethylene glycol, isopropanol, Tween-80, glycerol, propylene glycol or vegetable oil, and the vegetable oil is selected from soybean oil, castor oil, peanut oil, blended oil, etc.; the surfactant is selected from sodium dodecylbenzenesulfonate, stearic acid, polyoxyethylene-polyoxypropylene copolymer, sorbitan fatty acid ester or polysorbate (Tween), etc.; the flavoring agent is selected from aspartame, sucralose, essence, steviol glycoside, acesulfame potassium, citric acid or sodium saccharin; the preservative is selected from at least one of methyl paraben or propyl paraben.

[0093] In some embodiments of the present invention, the dosage form of the drug is selected from tablets (such as ordinary tablets, bilayer tablets, multilayer tablets, sustained-release tablets, single-chamber controlled-release tablets, double-chamber controlled-release tablets, microporous controlled-release tablets, sublingual tablets, orally disintegrating tablets, dispersible tablets, enteric-coated tablets), pills, powders, suspensions, gels, emulsions, creams, granules, nanoparticles, capsules (such as ordinary capsules, sustained-release capsules, controlled-release capsules, capsules containing pellets or tablets, pH-dependent capsules containing pellets or tablets, gastrointestinal complex capsules), suppositories, injections, sprays and injections.

[0094] The sixth aspect of the present invention provides the use of the water-soluble composite plant volatile oil solution of the first aspect of the present invention and / or the drug of the third aspect of the present invention in at least one of (1) to (8):

[0095] (1) Preparing a product for preventing and / or treating intestinal inflammation of meat ducks;

[0096] (2) Preparing a product for treating poor fecal form and diarrhea of meat ducks;

[0097] (3) Preparing a product for preventing and / or treating diseases caused by Escherichia coli infection in meat ducks;

[0098] (4) Preparing a product for preventing and / or treating diseases caused by Salmonella infection in meat ducks;

[0099] (5) Preparing a product for enhancing the disease resistance of the body of meat ducks;

[0100] (6) Prepare a product for repairing damaged mucosa of meat duck intestine;

[0101] (7) Prepare a product for improving the production performance and slaughter performance of meat ducks;

[0102] (8) Prepare an antibacterial product;

[0103] In some embodiments of the present invention, the antibacterial product in (8) has an antibacterial activity equivalent to that of antibiotics; the antibiotics include neomycin sulfate, apramycin, and colistin sulfate.

[0104] In the seventh aspect of the present invention, a product is provided, which includes the water-soluble composite plant volatile oil solution of the first aspect of the present invention or the drug of the third aspect of the present invention.

[0105] In some embodiments of the present invention, the product includes but is not limited to reagents, reagent kits, and drugs.

[0106] The beneficial effects of the present invention are:

[0107] The water-soluble composite plant volatile oil solution in the present invention has strong antibacterial activity, can replace antibiotics such as neomycin sulfate, apramycin, and colistin sulfate to effectively prevent and treat meat duck enteritis, has no withdrawal period, no drug residues, improves the clinical symptoms of loose feces, enhances intestinal health, ensures the slaughter rate, reduces the feed-to-meat ratio, increases the yield rate of duck gizzards, and improves the economic benefits of meat duck breeding and slaughter.

[0108] The present invention promotes the dissolution of volatile oils through modern processes and increases the yield of volatile oils. The high-pressure homogenization emulsification process is used in the preparation process to establish a nanoemulsion system, effectively improving the problem of poor water solubility of volatile oils. In addition, the present invention introduces oligosaccharide and vitamin components, which complement the advantages of volatile oil components, treating both the symptoms and the root causes. Administering the drug through drinking water can efficiently prevent and control and relieve the clinical enteritis of meat ducks, stabilize the intestinal tract of the duck flock, and ensure the breeding benefits of meat ducks. Description of the Drawings

[0109] Figure 1 Pictures of the appearance of the water-soluble composite plant volatile oil solution for meat ducks in Examples 1-14 and Comparative Examples 1-11.

[0110] Figure 2 HPLC chromatograms of cinnamaldehyde and carvacrol in the composite reference solution.

[0111] Figure 3 HPLC chromatograms of cinnamaldehyde and carvacrol in Example 1.

[0112] Figure 4 HPLC chromatograms of cinnamaldehyde and carvacrol in Example 2.

[0113] Figure 5It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 3.

[0114] Figure 6 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 4.

[0115] Figure 7 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 5.

[0116] Figure 8 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 6.

[0117] Figure 9 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 7.

[0118] Figure 10 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 8.

[0119] Figure 11 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 9.

[0120] Figure 12 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 10.

[0121] Figure 13 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 11.

[0122] Figure 14 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 12.

[0123] Figure 15 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 13.

[0124] Figure 16 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 14.

[0125] Figure 17 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 1.

[0126] Figure 18 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 2.

[0127] Figure 19 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 3.

[0128] Figure 20 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 4.

[0129] Figure 21It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 5.

[0130] Figure 22 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 6.

[0131] Figure 23 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 7.

[0132] Figure 24 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 8.

[0133] Figure 25 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 9.

[0134] Figure 26 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 10.

[0135] Figure 27 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 11.

[0136] Figure 28 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 1 at different accelerated times.

[0137] Figure 29 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Example 6 at different accelerated times.

[0138] Figure 30 It is the HPLC chromatogram of cinnamaldehyde and carvacrol in Comparative Example 8 at different accelerated times.

[0139] Figure 31 It is the enteritis feces state of the duck flock.

[0140] Figure 32 It is the result of intestinal pathological sections (HE staining) of each group before and after treatment. Detailed implementation manners

[0141] The concept of the present invention and the technical effects produced will be clearly and completely described below in conjunction with the embodiments to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0142] Example 1

[0143] A water-soluble compound plant volatile oil solution for meat ducks, by mass, includes 10 parts of cinnamon volatile oil, 9 parts of oregano volatile oil, 1 part of vitamin A, 0.1 part of mannan oligosaccharide, 35 parts of Tween-80, 10 parts of caprylic capric polyethylene glycol glyceride, 70 parts of diethylene glycol monoethyl ether, 0.3 part of potassium cinnamate, 0.2 part of butylated hydroxyanisole, 0.1 part of tea polyphenol, and 5 parts of water.

[0144] Among them, the preparation process of cinnamon volatile oil is as follows:

[0145] (1) Ultrafinely pulverize the dried cinnamon medicinal materials, pass through a 30-mesh sieve to obtain cinnamon pulverized matter;

[0146] (2) Moisten the cinnamon pulverized matter with 30% ethanol to control its water content at 25% to obtain moistened cinnamon medicinal materials;

[0147] (3) Then perform steam explosion on the moistened cinnamon medicinal materials at a pressure of 1.0 MPa, with a pressure maintenance time of 120 s and an explosion time of 0.01 s to obtain cinnamon steam explosion-treated matter;

[0148] (4) Subject the cinnamon steam explosion-treated matter to supercritical CO2 extraction, with an extraction pressure of 35 MPa, an extraction temperature of 40 ± 2 °C, an extraction time of 2.5 h, a CO2 flow rate of 18 L / h, and an extraction entrainer of ethanol with a concentration of 5%. Then obtain cinnamon volatile oil through low-temperature vacuum concentration.

[0149] In addition, the preparation process of oregano volatile oil is as follows:

[0150] (1) Ultrafinely pulverize the dried oregano medicinal materials, pass through a 40-mesh sieve to obtain oregano pulverized matter;

[0151] (2) Moisten the oregano pulverized matter with 30% ethanol to control its water content at 30% to obtain moistened oregano medicinal materials;

[0152] (3) Then perform steam explosion on the moistened oregano medicinal materials at a pressure of 0.8 MPa, with a pressure maintenance time of 100 s and an explosion time of 0.01 s to obtain oregano steam explosion-treated matter;

[0153] (4) Subject the oregano steam explosion-treated matter to supercritical CO2 extraction, with an extraction pressure of 20 MPa, an extraction temperature of 38 ± 2 °C, an extraction time of 1.5 h, a CO2 flow rate of 20 L / h, and an extraction entrainer of ethanol with a concentration of 9%. Then obtain oregano volatile oil through low-temperature vacuum concentration.

[0154] The preparation method of the above water-soluble compound plant volatile oil solution for meat ducks includes the following steps:

[0155] It includes 10 parts of cinnamon volatile oil, 9 parts of oregano volatile oil, 1 part of vitamin A, 0.1 part of mannan oligosaccharide, 35 parts of Tween-80, 10 parts of polyethylene glycol glycerol caprylate / caprate, 75 parts of diethylene glycol monoethyl ether, 0.3 part of potassium cinnamate, 0.2 part of butylated hydroxyanisole, 0.1 part of tea polyphenol, and 5 parts of water

[0156] By weight parts, mix vitamin A, mannan oligosaccharide, potassium cinnamate, butylated hydroxyanisole, tea polyphenol, polyethylene glycol glycerol caprylate / caprate, and diethylene glycol monoethyl ether by stirring to obtain Solution 1;

[0157] Mix Solution 1 with Tween-80 to obtain Solution 2;

[0158] Mix Solution 2 with cinnamon volatile oil and oregano volatile oil and emulsify. The emulsification pressure is 50 MPa, the temperature is 40 °C, and the time is 1 h to obtain the water-soluble composite plant volatile oil solution.

[0159] Example 2

[0160] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that it contains 7 parts of cinnamon volatile oil and 6 parts of oregano volatile oil.

[0161] Example 3

[0162] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that it contains 0.5 part of vitamin A and 0.05 part of mannan oligosaccharide.

[0163] Example 4

[0164] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that it contains 30 parts of Tween-80 and 75 parts of diethylene glycol monoethyl ether.

[0165] Example 5

[0166] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that the preservative is adjusted from potassium cinnamate to sodium benzoate with the same addition amount.

[0167] Example 6

[0168] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that the antioxidant is adjusted from butylated hydroxyanisole and tea polyphenol to only dibutylhydroxytoluene with the same addition amount.

[0169] Example 7

[0170] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 only in that the cinnamon medicinal materials are not subjected to steam explosion treatment during the preparation process of cinnamon volatile oil. The specific process is as follows:

[0171] (1) Accurately weigh the dried cinnamon medicinal materials and perform ultrafine pulverization. Pass through a 30-mesh sieve to obtain the pulverized cinnamon.

[0172] (2) Perform supercritical CO2 extraction on the pulverized cinnamon. The extraction pressure is 30 MPa, the extraction temperature is 40 ± 2 °C, the extraction time is 2.5 h, the CO2 flow rate is 18 L / h, the extraction entrainer is ethanol with a concentration of 5%. Then, perform low-temperature vacuum concentration to obtain cinnamon volatile oil.

[0173] Example 8

[0174] A water-soluble composite plant volatile oil solution for meat ducks, the difference from Example 1 is only that during the preparation process of oregano volatile oil, the oregano medicinal materials are not subjected to steam explosion treatment. The specific process is as follows:

[0175] (1) Accurately weigh the cinnamon and oregano medicinal materials and perform ultrafine pulverization. Pass through a 30-mesh sieve to obtain the pulverized oregano.

[0176] (2) Perform supercritical CO2 extraction on the pulverized oregano. The extraction pressure is 30 MPa, the extraction temperature is 40 ± 2 °C, the extraction time is 2.5 h, the CO2 flow rate is 18 L / h, the extraction entrainer is ethanol with a concentration of 5%. Then, perform low-temperature vacuum concentration to obtain oregano volatile oil.

[0177] Example 9

[0178] A water-soluble composite plant volatile oil solution for meat ducks, the difference from Example 1 is that no entrainer is used during the supercritical CO2 extraction process for both cinnamon volatile oil and oregano volatile oil.

[0179] Example 10

[0180] A water-soluble composite plant volatile oil solution for meat ducks, the difference from Example 1 is only that mannan oligosaccharide is replaced by xylo-oligosaccharide.

[0181] Example 11

[0182] A water-soluble composite plant volatile oil solution for meat ducks, the difference from Example 1 is that 5 parts of polyethylene glycol glycerol caprylate / caprate and 55 parts of diethylene glycol monoethyl ether are used.

[0183] Example 12

[0184] A water-soluble composite plant volatile oil solution for meat ducks, the difference from Example 1 is that the extraction pressure during the supercritical CO2 extraction process of cinnamon volatile oil is 20 MPa.

[0185] Example 13

[0186] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that the extraction pressure during the supercritical CO2 extraction of oregano volatile oil is 35 MPa.

[0187] Example 14

[0188] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that the emulsification pressure is 30 MPa.

[0189] Comparative Example 1

[0190] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that the extraction process of cinnamon volatile oil adopts the steam explosion process + steam distillation method, and the specific process is as follows:

[0191] (1) Accurately weigh the dried cinnamon medicinal materials and perform ultrafine pulverization, and pass through a 30-mesh sieve to obtain cinnamon pulverized matter;

[0192] (2) Moisten the cinnamon pulverized matter with 30% ethanol to control its water content at 25% to obtain moistened cinnamon medicinal materials;

[0193] (3) Then perform steam explosion on the moistened cinnamon medicinal materials at a pressure of 1.0 MPa, with a pressure maintenance time of 120 s and a blasting time of 0.01 s to obtain cinnamon steam explosion treated matter;

[0194] (4) Add 8 times the amount of water to the cinnamon steam explosion treated matter, heat it up to 95 ± 2 °C, distill for 3 h, and vacuum concentrate to an equal volume of cinnamon volatile oil.

[0195] Comparative Example 2

[0196] A water-soluble composite plant volatile oil solution for meat ducks, which is different from Example 1 in that the extraction process of oregano volatile oil adopts the steam explosion process + steam distillation method, and the specific process is as follows:

[0197] (1) Accurately weigh the dried oregano medicinal materials and perform ultrafine pulverization, and pass through a 40-mesh sieve to obtain oregano pulverized matter;

[0198] (2) Moisten the oregano pulverized matter with 30% ethanol to control its water content at 30% to obtain moistened oregano medicinal materials;

[0199] (3) Then perform steam explosion on the moistened oregano medicinal materials at a pressure of 0.8 MPa, with a pressure maintenance time of 100 s and a blasting time of 0.01 s to obtain oregano steam explosion treated matter;

[0200] (4) Add 8 times the amount of water to the oregano steam explosion treated matter, heat it up to 95 ± 2 °C, distill for 3 h, and vacuum concentrate to an equal volume of cinnamon volatile oil.

[0201] Comparative Example 3

[0202] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that the co-emulsifier is adjusted from polyethylene glycol glycerol caprylate / caprate and diethylene glycol monoethyl ether to an equal amount of water.

[0203] Comparative Example 4

[0204] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that the extraction pressure during the supercritical CO2 extraction of cinnamon volatile oil is 55 MPa.

[0205] Comparative Example 5

[0206] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that the extraction pressure during the supercritical CO2 extraction of oregano volatile oil is 45 MPa.

[0207] Comparative Example 6

[0208] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that the emulsification pressure is 55 MPa.

[0209] Comparative Example 7

[0210] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that no vitamins are added.

[0211] Comparative Example 8

[0212] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that no oligosaccharides are added.

[0213] Comparative Example 9

[0214] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that no antioxidants are added.

[0215] Comparative Example 10

[0216] A water-soluble composite plant volatile oil solution for meat ducks, which is different from that in Example 1 in that the extraction of oregano volatile oil is not carried out, and the oregano volatile oil in Example 1 is replaced with an equal amount of cinnamon volatile oil, that is: 19 parts of cinnamon volatile oil, 1 part of vitamin A, 0.1 part of mannan oligosaccharide, 35 parts of Tween-80, 10 parts of polyethylene glycol glycerol caprylate / caprate, 75 parts of diethylene glycol monoethyl ether, 0.3 part of potassium cinnamate, 0.2 part of butylated hydroxyanisole, 0.1 part of tea polyphenol, and 5 parts of water.

[0217] Comparative Example 11

[0218] A water-soluble compound plant volatile oil solution for meat ducks, which is different from Example 1 in that the extraction of cinnamon volatile oil is not carried out, and the cinnamon volatile oil in Example 1 is replaced with an equal amount of oregano volatile oil, namely: 19 parts of oregano volatile oil, 1 part of vitamin A, 0.1 part of mannan oligosaccharide, 35 parts of Tween-80, 10 parts of polyethylene glycol glycerol caprylate / caprate, 75 parts of diethylene glycol monoethyl ether, 0.3 part of potassium cinnamate, 0.2 part of butylated hydroxyanisole, 0.1 part of tea polyphenol, and 5 parts of water.

[0219] Effect Example

[0220] 1. Influence of different preparation conditions on the quality of the water-soluble compound plant volatile oil solution for meat ducks

[0221] 1.1 Detection items

[0222] 1.1.1 Characters

[0223] Observe the characters (color, clarity, presence of precipitation, turbidity, floccules, etc.) of the oral liquid and record (this product should be a light yellow to light brown clear liquid).

[0224] 1.1.2 Determination of the content of active ingredients by high performance liquid chromatography

[0225] (1) Determination of cinnamaldehyde content

[0226] Determine according to the high performance liquid chromatography method (Appendix 0512 of the Chinese Pharmacopoeia).

[0227] Chromatographic conditions and system suitability test: Use octadecylsilane chemically bonded silica gel as the filler, use acetonitrile-water (40:60) as the mobile phase, column temperature 35°C, flow rate 1.0 min / mL, injection volume 20 μL. The detection wavelength for cinnamaldehyde and carvacrol is 280 nm, and the number of theoretical plates calculated based on the cinnamaldehyde peak should not be less than ३०००.

[0228] Preparation of the stock solution of the cinnamaldehyde reference substance solution: Take 10 mg of cinnamaldehyde reference substance, accurately weigh it, place it in a 100 mL volumetric flask, add an appropriate amount of methanol to dissolve and dilute to the mark, and shake well. That is, the stock solution (1) is obtained.

[0229] Preparation of the mixed solution of carvacrol and cinnamaldehyde reference substances: Accurately weigh 5 mg of carvacrol and 1 mg of cinnamaldehyde respectively, place them in a 100 mL brown volumetric flask, dissolve with methanol and dilute to the mark, and shake well to prepare a mixed solution containing 50 μg / mL of carvacrol and 10 μg / mL of cinnamaldehyde.

[0230] Preparation of the test solution for the determination of thymol and carvacrol: Take 2.0 g of this product, place it in a 25 mL brown volumetric flask, add about 15 mL of methanol, shake well, add methanol to the mark, and shake well.

[0231] For the assay, 20 μL each of the reference substance solution and the test sample solution were accurately pipetted, injected into the liquid chromatograph, and determined to obtain the results.

[0232] The contents of cinnamaldehyde, thymol, and carvacrol in the samples of Examples 1 to 14 and Comparative Examples 1 to 11 were compared. The content calculation formula is as follows (during the calculation process, the content of the reference substance is calculated as 100%):

[0233]

[0234] 1.1.3 In vitro antibacterial evaluation

[0235] Among the intestinal pathogenic bacteria of meat ducks, Escherichia coli and Salmonella are relatively common. Therefore, the broth dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the water-soluble composite plant volatile oil solutions prepared in Examples 1 to 14 and Comparative Examples 1 to 11 against the Escherichia coli quality control strain (ATCC 25922) and the Salmonella quality control strain (CICC 10437). The positive control was Pulsatilla oral liquid (extracted according to the standards of the Pharmacopoeia of the People's Republic of China for Veterinary Drugs). First, each sample was diluted 8-fold with pure water. 100 μL of MH broth medium was added to columns 2 to 11 of the 96-well plate, 200 μL of MH broth medium was added to column 12 as a control, and 100 μL of each sample at 125 mg / mL was added to column 1. Using the two-fold dilution method, the concentrations in columns 1 to 11 were 62.5, 31.25, 15.63, 7.81, 3.90, 1.95, 0.98, 0.49, 0.24, 0.12, and 0 mg / mL, respectively, and the final concentration of the bacterial solution was 5×10 5 CFU / mL. The 96-well plate was placed in a biochemical incubator and cultured for 24 h. The lowest dilution concentration of the drug without obvious bacterial growth was the MIC (mg / mL) of the water-soluble composite plant volatile oil solution for this meat duck, and the lowest dilution concentration of the drug without bacterial growth after culturing on the drop plate for 24 h was the MBC (mg / mL) of the water-soluble composite plant volatile oil solution for this meat duck.

[0236] 1.2 Test results

[0237] 1.2.1 Results of appearance

[0238] The appearance results of Examples 1 to 14 and Comparative Examples 1 to 11 are shown in Table 1 and Figure 1 as follows. After using appropriate surfactants and co-emulsifiers, the appearance was basically uniformly transparent. In contrast, in Comparative Example 3, when the co-emulsifier was adjusted from diethylene glycol monoethyl ether and polyethylene glycol glycerol caprylate / caprate to water, a stable emulsion system could not be formed, resulting in stratification and turbidity. In addition, compared with Example 1, the solution system formed under an emulsification pressure of 55 Mpa in Comparative Example 5 also showed obvious demulsification, which would potentially affect the uniformity and functional effects of the active substances in the system.

[0239] Table 1 Properties of Examples and Comparative Examples

[0240]

[0241] 1.2.2 Content results

[0242] The contents of cinnamaldehyde and carvacrol in the water-soluble composite plant volatile oil solutions of the ducks of Examples 1 to 14 and Comparative Examples 1 to 11 were detected by HPLC. The test results are shown in Table 2. The HPLC spectrum of the standard solution is shown in Table 2. Figure 2 , wherein the elution time of cinnamaldehyde is 10.10min, and the elution time of carvacrol is 28.05min. The HPLC spectra of Examples 1 to 14 and Comparative Examples 1 to 11 are shown in Figures 3 to 27 .

[0243] Comparing Example 1 with Examples 3 to 6, Example 10, and Comparative Examples 6 to 8, the main difference lies in the addition of auxiliary materials such as preservatives, antioxidants, vitamins, and oligosaccharides. The test results show that there is no significant difference in the content of cinnamaldehyde and carvacrol (p>0.05), indicating that these ingredients do not affect the content of the active ingredients in the volatile oil.

[0244] Comparing the contents of cinnamaldehyde in Example 1 and Example 7, the results show that the pretreatment of cinnamon bark with steam explosion is more conducive to destroying the ultramicromolecular structure of cinnamon bark, promoting the dissolution of effective substances, and thus improving the extraction efficiency of steam explosion materials in supercritical extraction, and can significantly increase the content of cinnamaldehyde in cinnamon bark volatile oil by about 14.44% (p < 0.05).

[0245] Comparing the content of carvacrol in Example 1 and Example 8, the result trend is consistent with cinnamaldehyde. The pretreatment of oregano with steam explosion can significantly increase the content of carvacrol in oregano volatile oil by about 26.61% (p<0.05).

[0246] Comparing the contents of cinnamaldehyde and carvacrol in Example 1 and Example 9, the results showed that supercritical extraction using ethanol as an entrainer can enhance the extraction ability of cinnamaldehyde and carvacrol, and the contents of cinnamaldehyde and carvacrol increased by 17.58% (p<0.05) and 17.10% (p<0.05), respectively.

[0247] Comparing Example 1 with Comparative Examples 1-2, the supercritical CO2 extraction process significantly promoted the extraction efficiency of cinnamon volatile oil and oregano volatile oil compared with the steam distillation method, and the contents of cinnamaldehyde and carvacrol were increased by 31.58% (p<0.05) and 87.96% (p<0.05), respectively.

[0248] Comparing Example 1 and Comparative Example 3, after the co-emulsifier was replaced by water, a stable emulsified system could not be formed, and stratification occurred. In the upper emulsion, the contents of cinnamaldehyde and carvacrol were significantly lower than those in Example 1, decreasing by 56.10% (p<0.05) and 55.59% (p<0.05), respectively.

[0249] Comparing Example 1, Example 12 and Comparative Example 4, the results show that the cinnamaldehyde content of Example 1 and Example 12 is similar, with no significant difference (p>0.05), but is significantly better than Comparative Example 4, and its cinnamaldehyde content decreases by 31.13% (p<0.05) compared with Example 1, indicating that the supercritical extraction pressure of cinnamon volatile oil is better than 55Mpa between 20~35Mpa, and too high extraction pressure cannot promote the effective dissolution of cinnamaldehyde, and actual operation should be controlled within an appropriate extraction pressure range.

[0250] Comparing Example 1, Example 13 and Comparative Example 5, the results show that the carvacrol content of Example 1 and Example 13 is similar, with no significant difference (p>0.05), but is significantly better than Comparative Example 5, and its carvacrol content is reduced by 44.56% (p<0.05) compared with Example 1, indicating that the supercritical extraction pressure of oregano volatile oil is better than 45 MPa when it is between 20 and 35 MPa, and that too high an extraction pressure cannot promote the effective dissolution of carvacrol.

[0251] Comparing Example 1, Example 14 and Comparative Example 5, the results show that when the emulsification pressure is between 30 and 50 MPa, there is no significant difference in the content of cinnamaldehyde and carvacrol in Example 1 and Example 14 (p>0.05). However, when the emulsification pressure is increased to 55 MPa, the solution becomes turbid from clear, and the content of cinnamaldehyde and carvacrol are significantly decreased by 19.62% (p<0.05) and 19.67% (p<0.05) compared with Example 1, indicating that increasing the emulsification pressure will destroy the emulsification equilibrium of the system and cause demulsification.

[0252] Table 2 Cinnamaldehyde and carvacrol contents in Examples and Comparative Examples

[0253]

[0254]

[0255] Note: In Table 2, a represents that the cinnamaldehyde content in the sample is significantly different from that in Example 1, p<0.05; b represents that the carvacrol content in the sample is significantly different from that in Example 1, p<0.05.

[0256] 1.2.3 In vitro antibacterial evaluation results

[0257] The in vitro antibacterial (MIC and MBC) assay results of Examples 1-14 and Comparative Examples 1-11 against Escherichia coli, Salmonella, and Clostridium perfringens are shown in Table 3. The antibacterial activities of Examples 1-14 and Comparative Examples 1-11 against Escherichia coli, Salmonella, and Clostridium perfringens are significantly stronger than those of Pulsatilla root oral liquid, with MIC < 7.80 mg / mL and MBC < 15.60 mg / mL.

[0258] Comparing Example 1 with Examples 7-9 and Comparative Examples 1-5, Example 1 has the best antibacterial activity, corresponding to the highest contents of cinnamaldehyde and carvacrol in 1.2.2, indicating that the overall antibacterial activity trend is positively correlated with the content of active ingredients. The steam explosion technology combined with supercritical CO2 extraction mode is the optimal extraction mode.

[0259] Comparing Example 1 with Comparative Examples 10-11, Example 1 has better antibacterial activity, indicating that the combined antibacterial activity of cinnamon volatile oil and oregano volatile oil is better than that of a single volatile oil under the same number of parts.

[0260] Table 3 Comparison results of in vitro antibacterial activities of Examples 1-15 and Comparative Examples 1-11

[0261]

[0262] Note: The unit of MIC is mg / mL.

[0263] 2. Improvement effect of different antioxidants on the stability of active ingredients in cinnamon volatile oil / oregano volatile oil

[0264] Considering that the core volatile oil components of the present invention, cinnamaldehyde and carvacrol, may be oxidized and decomposed, especially since the cinnamaldehyde molecule contains unsaturated chemical bonds and is extremely easy to be oxidized and decomposed into components such as cinnamic acid, reducing the antibacterial activity. Therefore, antioxidants were added to try to maintain the stability of the system. In the present invention, the effects of the conventional antioxidant dibutylhydroxytoluene (Example 6), the combination of butylated hydroxyanisole + tea polyphenols (Example 1), and no additional antioxidant (Comparative Example 9) on the stability of cinnamaldehyde were mainly evaluated. All samples were prepared according to the preparation method of Example 1.

[0265] 2.1 Stability investigation conditions

[0266] The above Examples 1, 6, and Comparative Example 9 were placed in a stability test chamber. According to the requirements of the stability research technical guidelines in the "Compilation of Technical Guidelines for Veterinary Drug Research", the product properties and the contents of cinnamaldehyde and carvacrol were subjected to accelerated investigation. The accelerated conditions were (40 ± 2) °C and relative humidity (75 ± 5)%. Samples were taken for testing at 0, 3, and 6 months respectively to investigate the properties of the samples and the content of cinnamaldehyde, analyze the change trend of cinnamaldehyde, and determine a more suitable antioxidant.

[0267] 2.2 Test results

[0268] The HPLC chromatograms of the contents of cinnamaldehyde and carvacrol in Example 1, Example 6, and Comparative Example 9 after different acceleration times are shown in Figures 28 to 30 .

[0269] The content results are shown in Table 4. Comparing Example 1, Example 6, and Comparative Example 9, the results show that adding antioxidants can significantly inhibit the oxidative degradation of cinnamaldehyde and carvacrol in the formulation of the present invention. In terms of the selection of antioxidants, the combination antioxidant composition of butylated hydroxyanisole + tea polyphenols in Example 1 has a better effect than using only dibutylhydroxytoluene in Example 6. Among them, the degradation rate of cinnamaldehyde after 6 months of acceleration is only 0.96%, and the degradation rate of carvacrol is only 1.79%, which is significantly better than that of Example 6 (p < 0.05) and Comparative Example 9 (p < 0.05).

[0270] Table 4 Results of stability investigation

[0271]

[0272] Note: In Table 4, a represents a significant difference in the content of cinnamaldehyde in the sample compared with Example 1 in the same period, p < 0.05; b represents a significant difference in the content of carvacrol in the sample compared with Example 1 in the same period, p < 0.05.

[0273] 3. Evaluation of the preventive effect of enteritis in meat ducks

[0274] 3.1 Test method

[0275] 3.1.1 Test modeling and grouping

[0276] Select 200 6-day-old Cherry Valley meat ducks with similar body weights and good health, and randomly divide them into 10 groups, including 1 blank group, 1 model group, and 8 experimental groups. The specific treatment methods are as follows: (1) Blank group: No challenge is carried out, and no drug treatment is used; (2) Model group: Salmonella is intragastrically administered orally at a dose of 1×10 9 cfu / duck, and no drug treatment is used; (3) Experimental groups 1-4: Each group of meat ducks is first intragastrically administered orally with Salmonella at a dose of 1×10 9cfu / feather. Three days before virus challenge, different drugs were used for prevention. The drugs corresponding to Example 1, Example 2, Example 3, and Example 10 were used for prevention respectively. The administration method was to mix the drugs with water at a ratio of 1:1000 and administer the drugs for 8 hours every day. (4) For experimental groups 5 - 6, the drugs corresponding to Comparative Example 7 and Comparative Example 8 were used for prevention respectively. (5) For the positive control group 1, Radix Pulsatillae Oral Liquid was used for prevention, with a dosage of 0.5 mL per duck per day. (6) For the positive control group 2, the natural plant feed additive in the authorized patent document CN102948641B was used for prevention, and it was added to the feed at a ratio of 0.15%. The virus challenge and administration methods for experimental groups 5 - 8 were the same as those for experimental groups 1 - 4.

[0277] 3.1.2 Efficacy evaluation

[0278] (1) Statistical analysis of prevention efficiency: Count the number of meat ducks showing diarrhea symptoms from each group after virus challenge until 10 - day - old, and calculate the prevention efficiency.

[0279] (2) Statistical analysis of mortality rate: Count the total number of deaths in each group until 10 - day - old, and calculate the mortality rate.

[0280] 3.2 Test results

[0281] By establishing a prevention - virus challenge model, the diarrhea rates and survival rates of each group were finally counted as shown in Table 5. The model group showed that about 90% of the meat ducks had diarrhea after virus challenge, and about 30% mortality rate occurred during the same period, indicating that the virus challenge with this concentration of Salmonella could effectively cause enteritis in meat ducks, and even death occurred without intervention treatment. Compared with the model group, each experimental group could reduce the diarrhea rate and mortality rate of the duck flock to a certain extent. The comparison results between the experimental groups showed that the examples and comparative examples involved in the present invention were superior to the positive control group of Radix Pulsatillae Oral Liquid and the natural plant feed additive in the authorized patent document CN102948641B in preventing Salmonella diarrhea. In particular, for Example 1, Example 2, Example 3, and Example 10, the diarrhea rate was reduced by more than 25% compared with the positive control group, and the mortality rate was reduced by more than 20%.

[0282] Comparing experimental group 1 and experimental group 2, the mortality rates were the same. In terms of controlling the diarrhea rate, experimental group 1 was superior to experimental group 2, indicating that the preventive effect was dose - dependent on the contents of cinnamaldehyde and carvacrol, and it was reflected that the antibacterial activity was particularly crucial for the preventive effect.

[0283] Comparing experimental group 1, experimental group 3, and experimental group 5, in terms of mortality rate: experimental group 5 > experimental group 3 = experimental group 1; in terms of diarrhea rate: experimental group 5 > experimental group 3 > experimental group 1, indicating that the preventive effect was dose - dependent on the content of vitamin A, and the addition of vitamin A showed a potential protective effect on the intestinal mucosa.

[0284] For Comparative Test Group 1, Test Group 4, and Test Group 6, in terms of mortality rate: Test Group 6 > Test Group 4 = Test Group 1; in terms of diarrhea rate: Test Group 6 > Test Group 4 > Test Group 1. This indicates that when preventing and controlling bacterial enteritis, adding oligosaccharides has an obvious synergistic effect, and mannan oligosaccharide is superior to xylo-oligosaccharide.

[0285] Table 5 Statistics of Diarrhea and Death in the Prevention-Challenge Model

[0286]

[0287]

[0288] 4. Evaluation of the Therapeutic Effect of Duck Enteritis

[0289] 4.1 Test Subjects

[0290] 31-day-old meat ducks from a certain meat duck breeding company in Shandong Province, with a single-house inventory of 12,000 ducks. Affected by the high temperature and high humidity in summer, the duck flock showed symptoms of loose stools and diarrhea (see Figure 31 ), which are typical clinical symptoms of bacterial enteritis. To verify the therapeutic effect of this product on diarrhea caused by bacterial enteritis, 350 meat ducks with pasted anus were selected, with their body weights controlled at 2060 ± 10 g. They were evenly divided into 7 groups, with 50 ducks in each group, namely the blank control group (without drug treatment), Test Group 1 (treated with Example 1, and the drug was diluted in water at a ratio of 1:1000 and administered for 8 hours every day), Test Group 2 (treated with Patrinia oral liquid, administered orally at 0.5 mL / duck / day), Test Group 3 (treated with the drug described in the authorized patent CN102948641B, added to the feed at 0.15%), Test Groups 4 - 6 (treated with 32.5% neomycin sulfate, 10% apramycin sulfate, and colistin sulfate respectively according to the medication guidance). The drugs were continuously administered for 5 days (the administration time corresponded to 32 - 36 days of age) for efficacy evaluation.

[0291] 4.2 Efficacy Evaluation Indicators

[0292] 4.2.1 Feed Intake Statistics

[0293] During the test period, the duck flocks in each group maintained the feeding habit of free access to feed and water. The total daily feed intake of each day from 31 to 37 days of age was continuously monitored, and the average daily feed intake per duck was calculated using the following formula:

[0294] Average daily feed intake per duck = Total daily feed intake ÷ Number of surviving ducks in each group.

[0295] 4.2.2 Monitoring of Fecal Improvement

[0296] The statistics of loose feces in the house and the pasted anus rate were carried out on days 31, 34, and 37 respectively.

[0297] The method for thin dung statistics is as follows: Enter from the entrance of the duck house, walk towards the center, randomly take 20 samples of fresh dung, and conduct thin dung rate statistics:

[0298] Thin dung rate = number of thin dung ÷ 20 × 100%.

[0299] The method for pasty anus rate statistics is as follows: Randomly select 30 ducks from each group to observe the condition of pasty anus, record the number of ducks with pasty anus, and calculate the pasty anus rate:

[0300] Pasty anus rate = number of pasty anus ÷ 30 × 100%.

[0301] 4.2.3 Growth performance statistics

[0302] Randomly select 30 ducks before feeding at 31 days old and after feeding at 37 days old respectively for body weight measurement, record the average body weight, total weight gain and average daily weight gain of each group, and compare the changes in production performance of each group after the occurrence of enteritis.

[0303] Average daily weight gain = (average body weight (37日龄) - average body weight (31日龄) ) ÷ 7.

[0304] 4.2.4 Mortality and culling statistics

[0305] Record the daily mortality and culling numbers of each group, count the total mortality and culling numbers of each group within 31 - 37 days old, and calculate the mortality and culling rate:

[0306] Mortality and culling rate = total mortality and culling numbers ÷ 50 × 100%

[0307] 4.2.5 Intestinal morphology observation

[0308] Before grouping, select diarrheal ducks for dissection, take their jejunum segments. After the experiment, complete the above data recording and then randomly select 3 meat ducks from each group for dissection, take their jejunum segments, and conduct intestinal tissue morphology observation by HE staining method to evaluate the recovery effect.

[0309] 4.3 Test results

[0310] 4.3.1 Feed intake statistics

[0311] The statistics of average daily feed intake per duck during the test are shown in Table 6. As the age of the duck group increases, the feed intake of each group shows a regular increasing trend. At the same age, there is no significant difference in feed intake among the experimental groups (P > 0.05), which proves that the drugs in each group will not affect the feed intake of meat ducks.

[0312] Table 6 Feed intake statistics table (g / duck / day)

[0313]

[0314] 4.3.2 Fecal improvement monitoring

[0315] Before drug administration (at 31 days old), during drug administration (at 34 days old), and after drug administration (at 37 days old), the loose feces and pasted anus conditions of each group of ducks were monitored and statistically analyzed. The statistical results are shown in Table 7. Before treatment, the loose feces rate of each group was in the state of a large proportion of loose feces at 75% - 90%. After 3 days of drug treatment, each group had a certain degree of relief. The loose feces rate: Group 1 < Group 4 = Group 6 < Group 5 < Group 3 < Group 2 < Blank control group. The improvement effect of Group 1 was the most obvious. The same trend was also reflected in the statistics at 37 days old, indicating that the present invention is superior to antibiotic treatment in improving loose feces.

[0316] In the statistics of the pasted anus rate, drug treatment was started when the pasted anus rate of each group was close to 100%. After 5 days, the pasted anus rate: Group 1 < Group 6 < Group 5 < Group 4 < Group 2 < Group 3, indicating that the present invention has the best effect in treating pasted anus.

[0317] Table 7 Monitoring of feces and pasted anus conditions

[0318]

[0319]

[0320] 4.3.3 Growth performance statistics

[0321] The statistical results of growth performance during the experiment are shown in Table 8. The initial average weight of each group was close, about 2060 ± 10 g / duck. As the disease process developed, the growth of the blank control group was significantly inhibited due to no drug intervention. Under the current breeding mode, the standard weight of healthy meat ducks at 37 days old should be close to 2802 g / duck, while the average weight of the blank control group at 37 days old was only 2404.04 g / duck, indicating that the serious diarrhea problem had a great impact on the feed conversion process. In contrast, each experimental group had a certain effect during the treatment. The ranking of the average weight at 37 days old: Group 1 > Group 5 > Group 4 > Group 6 > Group 2 > Group 3 > Blank control group. Among them, Group 1 had the best effect, with an average weight of 2797.17 g, significantly higher than the blank control group (p < 0.05). The effects of the antibiotic treatment groups (Groups 4 - 6) were similar, and the final average weight was between 2660 - 2670 g, but still lower than that of Group 1. The weights of Groups 2 and 3 were better than that of the blank control group, but the recovery effect was limited, mainly probably due to their weaker antibacterial activity.

[0322] Each experimental group showed the same trend as the final weight in the statistics of total weight gain and daily weight gain. Group 1 showed the best recovery effect during the treatment process, indicating that the present invention can play a better effect compared with traditional antibiotic treatment, promote the recovery of the production performance of ducks, and improve the feed conversion rate.

[0323] Statistical results of growth performance during the test period in Table 8

[0324]

[0325] Note: In Table 8, a represents that the average body weight of the test group has a significant difference compared with that of the control group in the same period, p < 0.05; b represents that the total weight gain of the test group has a significant difference compared with that of the control group, p < 0.05, bb represents that the total weight gain of the test group has a highly significant difference compared with that of the control group, p < 0.01; cc represents that the average daily weight gain of the test group has a highly significant difference compared with that of the control group, p < 0.01.

[0326] 4.3.4 Mortality and culling statistics

[0327] The daily mortality and culling numbers of each group during the test period were counted, and the results are shown in Table 9. The total mortality and culling numbers: Group 1 < Group 4 < Group 5 = Group 6 < Group 2 = Group 3 < Blank control group. This result shows that on the one hand, if the serious diarrhea problem is not intervened and treated in time, it will cause a high mortality rate. On the other hand, it shows that Group 1, as a natural medicine preparation, is superior to the traditional Chinese medicine preparation - Patrinia decoction and other Chinese medicine prescriptions in controlling mortality and culling. Although antibiotics also play a strong curative effect on controlling mortality and culling, the mortality and culling rate is slightly higher than that of Group 1, which once again reflects that Group 1 has the unique astringent effect of traditional Chinese medicine in addition to the high antibacterial activity similar to antibiotics.

[0328] Table 9 Statistical results of mortality and culling during the test period

[0329]

[0330] 4.3.5 Intestinal tissue morphology observation

[0331] After dissecting the duck flocks before and after treatment, small intestine pathological sections were intercepted for staining and microscopic examination. The results are shown in Figure 32 , and it can be observed that before treatment, there was obvious epithelial cell shedding in the small intestine, and the intestinal villi were short, indicating the occurrence of enteritis;

[0332] After 5 days of treatment, there was a large amount of intestinal epithelial cell shedding in the small intestine of the blank control group, the intestinal villi were fragmented and disappeared, the lamina propria was exposed, the crypts disappeared, and the intestinal absorption function was lost; while the 5 test groups all had varying degrees of recovery. Among them, Group 1 had the best recovery effect, with only a small amount of epithelial cell shedding in the intestine, relatively complete villi, obvious crypts, and no obvious inflammatory changes; followed by Groups 2 - 3 with partial intact intestinal villi and a small amount of intestinal epithelial cell necrosis; the worst effect was in Groups 4 - 6, where only part of the intestinal villi recovered, the crypts were not obvious, and there was still a large amount of intestinal epithelial cell shedding and necrosis.

[0333] 5. Influence of whole-cycle enteritis health care of meat ducks on production performance and slaughter performance

[0334] 5.1 Test subjects

[0335] A certain meat duck breeding company in Shandong Province has a total inventory of 150,000 ducks. 600 ducks are randomly selected and divided into 6 groups for a comparative test. Among them, 3 groups use 32.5% neomycin sulfate for whole-course health care as control groups 1 - 3, and the other 3 groups use Example 1 of the present invention for whole-course health care as test groups 1 - 3 to evaluate the final production performance and slaughter performance.

[0336] 5.2 Evaluation indicators

[0337] 5.2.1 Production performance

[0338] All duck flocks are sold at 38 days of age. Independent data statistics are carried out for the control group and the test group respectively, including three key indicators: selling weight, survival rate, and feed-to-meat ratio.

[0339] 5.2.2 Slaughter performance

[0340] Follow up the slaughter of each group of duck flocks. Randomly weigh 50 duck gizzards and duck intestines in each group to calculate the average weight and evaluate the influence of the present invention on the weight of duck gizzards and duck intestines compared with antibiotics.

[0341] 5.3 Test results

[0342] 5.3.1 Statistics of production performance

[0343] The statistical results of the production performance of each group are shown in Table 10. The average selling weight of control groups 1 - 3 is 2883.14 - 2905.52 g / duck, and the average selling weight of test groups 1 - 3 is 2992.58 - 3012.82 g / duck, with an average increase of 90.91 g / duck; the selling survival rate of control groups 1 - 3 is 96% - 98%, and the selling survival rate of test groups 1 - 3 is 98% - 100%, with an average increase of 2%; the feed-to-meat ratio of control groups 1 - 3 is 1.72 - 1.73, and the feed-to-meat ratio of test groups 1 - 3 is 1.67 - 1.69, with an average decrease of 0.04. It shows that the present invention can improve the average selling weight and survival rate of duck flocks and reduce the feed-to-meat ratio compared with traditional antibiotics for intestinal health care, greatly improving the economic benefits of terminal breeding and realizing the value of antibiotic substitution.

[0344] Table 10 Statistics of production performance

[0345]

[0346] 5.3.1 Statistics of slaughter performance

[0347] The present invention has established the exact effect on the prevention and control of enteritis in meat ducks, and further evaluated the influence on the weights of the gizzards and duck intestines in the digestive system. The results are statistically shown in Table 11, indicating that the weights of the gizzards in Control Groups 1 - 3 are 51.80 - 53.21 g / pair, while the weights of the gizzards in Experimental Groups 1 - 3 are 55.91 - 57.09 g / pair, with an average increase of 4.05 g / pair; the weights of the duck intestines in Control Groups 1 - 3 are 95.36 - 99.70 g / pair, and the weights of the duck intestines in Experimental Groups 1 - 3 are 101.67 - 105.18 g / pair, with an average increase of 5.56 g / pair. The results demonstrate that the present invention can promote organ development to a certain extent, increase intestinal integrity, and ultimately be reflected in the weights of the gizzards and duck intestines by regulating the intestinal tract of meat ducks and accelerating nutrient absorption.

[0348] Table 11 Statistical Results of Slaughter Performance

[0349]

[0350] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the scope of knowledge possessed by those of ordinary skill in the relevant art. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A water-soluble composite plant volatile oil solution, comprising the following components: cinnamon volatile oil, oregano volatile oil, vitamins, oligosaccharides, surfactants, co-emulsifiers, preservatives and antioxidants.

2. The water-soluble composite plant volatile oil solution according to claim 1, wherein: By mass, the water-soluble composite plant volatile oil solution comprises 7-12 parts of cinnamon volatile oil, 6-10 parts of oregano volatile oil, 0.5-1.5 parts of vitamins, 0.01-0.1 parts of oligosaccharides, 30-45 parts of surfactants, 60-85 parts of co-emulsifiers, 0.005-0.4 parts of preservatives and 0.01-0.5 parts of antioxidants.

3. The water-soluble composite plant volatile oil solution according to claim 1, wherein: The vitamins include at least one of vitamin A, vitamin D3, vitamin E, vitamin K3; and / or The oligosaccharides include one of mannan oligosaccharide and xylo-oligosaccharide; and / or The surfactants include at least one of Tween-40, Tween-60, Tween-80; and / or The co-emulsifiers include at least one of ethanol, glycerol, propylene glycol monocaprylate, polyethylene glycol glycerol caprylate / caprate, diethylene glycol monoethyl ether; and / or The antioxidants include at least one of butylated hydroxyanisole, dibutylhydroxytoluene, tea polyphenols, tert-butylhydroquinone; and / or The preservatives include at least one of potassium cinnamate, sodium benzoate, ethylparaben.

4. The water-soluble composite plant volatile oil solution according to any one of claims 1 to 3, wherein: The cinnamon volatile oil is prepared by the following method: Crush the dried cinnamon medicinal material, sieve it through a mesh sieve to obtain a cinnamon crushed material; Infiltrate the cinnamon crushed material with ethanol to obtain an infiltrated cinnamon medicinal material; Then perform steam explosion on the infiltrated cinnamon medicinal material to obtain a cinnamon steam explosion treated material; Use the supercritical CO2 extraction method to extract and concentrate the volatile oil from the cinnamon steam explosion treated material to obtain the cinnamon volatile oil; Preferably, the pressure of the steam explosion is 0.6-1.5 MPa, and the pressure maintenance time is 60-240 s; Preferably, the conditions for supercritical CO2 extraction are an extraction pressure of 15-50 MPa, an extraction temperature of 30-60 °C, an extraction time of 1-4 h, and a CO2 flow rate of 5-30 L / h; Preferably, an entrainer is added in the supercritical CO2 extraction, and the entrainer includes one of methanol, ethanol, and acetone, and the addition ratio of the entrainer is 1%-8%.

5. The water-soluble composite plant volatile oil solution according to any one of claims 1 to 3, wherein: The oregano volatile oil is prepared by the following method: Crush the dried oregano medicinal material, sieve it through a mesh sieve to obtain a pretreated oregano medicinal material; Infiltrate the pretreated oregano medicinal material with ethanol to obtain an infiltrated oregano medicinal material; Then perform steam explosion on the infiltrated oregano medicinal material to obtain an oregano steam explosion treated material; Use the supercritical CO2 extraction method to extract and concentrate the volatile oil from the oregano steam explosion treated material to obtain the oregano volatile oil; Preferably, the pressure of the steam explosion is 0.4-1.2 MPa, and the pressure maintenance time is 60-180 s; Preferably, the conditions for supercritical CO2 extraction are as follows: extraction pressure 8 - 40 MPa, extraction temperature 35 - 50 °C, extraction time 1 - 3 h, and CO2 flow rate 10 - 35 L / h; Preferably, an entrainer is added in the supercritical CO2 extraction. The entrainer includes one of methanol, ethanol, and acetone, and the addition ratio of the entrainer is 3% - 10%.

6. The method for preparing the water-soluble cinnamon volatile oil solution according to any one of claims 1 - 5, comprising the following steps: Stir and mix vitamins, oligosaccharides, preservatives, antioxidants, and co-emulsifiers to obtain Solution 1; Mix Solution 1 with a surfactant to obtain Solution 2; Mix Solution 2 with cinnamon volatile oil and oregano volatile oil and emulsify to obtain a water-soluble composite plant volatile oil solution.

7. According to the preparation method described in claim 6, it is characterized in that: The conditions for emulsification are a pressure of 25 MPa - 50 MPa; and / or The temperature is 25 °C - 50 °C, and / or The emulsification time is 0.25 - 1.5 h.

8. A drug comprising the water-soluble composite plant volatile oil solution according to any one of claims 1 - 5.

9. Application of the water-soluble composite plant volatile oil solution according to any one of claims 1 - 5 and / or the drug according to claim 8 in at least one of (1) - (8): (1) Preparing a product for preventing and / or treating intestinal inflammation in meat ducks; (2) Preparing a product for treating poor fecal form and diarrhea in meat ducks; (3) Preparing a product for preventing and / or treating diseases caused by Escherichia coli infection in meat ducks; (4) Preparing a product for preventing and / or treating diseases caused by Salmonella infection in meat ducks; (5) Preparing a product for enhancing the disease resistance of the body of meat ducks; (6) Preparing a product for repairing damaged intestinal mucosa in meat ducks; (7) Preparing a product for improving the production performance and slaughter performance of meat ducks; (8) Preparing an antibacterial product.

10. A product comprising the water-soluble composite plant volatile oil solution according to any one of claims 1 - 5 or the drug according to claim 8.

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